Project description:In this study the transcriptomes of Acinetobacter baumannii strains ATCC 17978 and 17978hm were compared. Strain 17978hm is a hns knockout derivative of strain ATCC 17978. Strain 17978hm displays a hyper-motile phenotype on semi-solid Mueller-Hinton (MH) media (0.25% agar). ATCC 17978 and 17978hm from an 37C overnight culture were transferred to the centre of the semi-solid MH plate and incubated at 37C for 8 hours. Only 17978hm cells displayed a motile phenotype and covered the complete surface of the plate. These motile 17978hm cells and the non-motile wild-type ATCC 17978 cells were harvested and RNA was isolated. The comparative transcriptome analysis was performed using the FairPlay labeling kit and a custom made Agilent MicroArray with probes designed to coding regions of the ATCC 17978 genome. The data was analyzed using Agilent GeneSpring GX9 and the significance analysis of microarray MS Excel add-on.
Project description:To further elucidate the mechanism of BSP-II on immunity on the broad molecular level, we have employed whole genome microarray expression profiling as a discovery platform to examine gene expression patterns during BSP-II treatment in a mouse derived hybridoma culture system. Hybridoma cell was treated ex vivo, and robust normalization of the data identified 1279 differentially expressed probe sets exhibiting minimum 1.5-fold changes that distinguished between BSP-II and control samples. Various pathwayswere significantly impacted by BSP-II treatment, and gene Ontology annotations show changes in the expression of molecules involved in immune and immunocyte related cellular processes. Expression of nine genes (MS4A2, CD3D, FGF21, CD80, PTPRC, NFATC4, IL2RB, Fas and LAT) from this signature was quantified in the RNA samples by QRT-PCR, confirming low variability between the predicted response patterns.
Project description:To further elucidate the mechanism of BSP-II on immunity on the broad molecular level, we have employed whole genome microarray expression profiling as a discovery platform to examine gene expression patterns during BSP-II treatment in a mouse derived hybridoma culture system. Hybridoma cell was treated ex vivo, and robust normalization of the data identified 1279 differentially expressed probe sets exhibiting minimum 1.5-fold changes that distinguished between BSP-II and control samples. Various pathwayswere significantly impacted by BSP-II treatment, and gene Ontology annotations show changes in the expression of molecules involved in immune and immunocyte related cellular processes. Expression of nine genes (MS4A2, CD3D, FGF21, CD80, PTPRC, NFATC4, IL2RB, Fas and LAT) from this signature was quantified in the RNA samples by QRT-PCR, confirming low variability between the predicted response patterns. BSP-II induced gene expression in hybridoma cell was measured at 4 hours after exposure to doses of 0ug/ml and 5ug/ml. Two independent experiments were performed using different cells for each experiment.
